In vitro tests for the measurement of clostridial toxins, toxoids and antisera. II. Titration of Clostridium perfringens toxins and antitoxins in cell culture.
The usefulness of cytopathic indicators for the titration of Cl perfringens beta and epsilon toxins has been investigated. Neutralization experiments with monoclonal antibodies have shown that the entities responsible for the lethal and dermonecrotic effects of Cl perfringens beta toxin preparations are identical. However, the cytopathic effects of the same preparations are caused by other entities. Therefore, titrations based upon lethal and dermonecrotic indicators of beta toxin are equally valid but those based on cytopathic effects are not. Similar experiments with Cl perfringens epsilon preparations have shown that their lethal, dermonecrotic and cytopathic activities are all caused by the same entity. It follows that all three activities can be valid indicators for toxin neutralization tests. Cell culture titrations of Cl perfringens epsilon antitoxin performed on rabbit sera at the levels of test prescribed by the European Pharmacopoeia have produced consistent results which agree closely with the dermonecrotic test. This test has, in turn, been shown to reflect the results of the mouse lethal test accurately. Titrations of cattle and sheep sera at lower levels of test have also produced results in close agreement with the in vivo test. It is concluded that cell culture titration offers a valid in vitro alternative to the use of mouse lethal and guinea-pig dermonecrotic indicators for the titration of sera generated in the course of potency tests and field trials of Cl perfringens epsilon vaccines.
Knight
P A
PA
Wellcome Biotechnology Ltd, Beckenham, Kent, U.K.
Queminet
J
J
Blanchard
J H
JH
Tilleray
J H
JH
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins / Q000032:analysis* / Q000276:immunology; D001428:Bacterial Vaccines / Q000032:analysis; D001681:Biological Assay / Q000379:methods; D002460:Cell Line; D003016:Clostridium perfringens / Q000032:analysis* / Q000276:immunology; D009500:Neutralization Tests; D014121:Toxoids / Q000032:analysis*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; C035454:Clostridium perfringens toxoid; D014121:Toxoids